Abstract
As a tightly controlled biological process, cardiogenesis requires the specification and migration of a suite of cell types to form a particular three-dimensional configuration of the heart. Many genetic factors are involved in the formation and maturation of the heart, and any genetic mutations may result in severe cardiac failures. The neuron navigator (NAV) family consists of three vertebrate homologs (NAV1, NAV2, and NAV3) of the neural guidance molecule uncoordinated-53 (UNC-53) in Caenorhabditis elegans. Although they are recognized as neural regulators, their expressions are also detected in many organs, including the heart, kidney, and liver. However, the functions of NAVs, regardless of neural guidance, remain largely unexplored. In our study, we found that nav3 gene was expressed in the cardiac region of zebrafish embryos from 24 to 48 h post-fertilization (hpf) by means of in situ hybridization (ISH) assay. A CRISPR/Cas9-based genome editing method was utilized to delete the nav3 gene in zebrafish and loss of function of Nav3 resulted in a severe deficiency in its cardiac morphology and structure. The similar phenotypic defects of the knockout mutants could recur by nav3 morpholino injection and be rescued by nav3 mRNA injection. Dual-color fluorescence imaging of ventricle and atrium markers further confirmed the disruption of the heart development in nav3-deleted mutants. Although the heart rate was not affected by the deletion of nav3, the heartbeat intensity was decreased in the mutants. All these findings indicate that Nav3 was required for cardiogenesis in developing zebrafish embryos.
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Data Availability
All data generated or analyzed during this study are included in this published article and its supplementary information files.
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Abbreviations
- CH:
-
Calponin homology
- dpf:
-
Days post-fertilization
- HI:
-
Heartbeat intensity
- hpf:
-
Hours post-fertilization
- ISH:
-
In situ hybridization
- NAV:
-
Neuron navigator
- SH3:
-
Src homology 3
- UNC53:
-
Uncoordianted-53
- WISH:
-
Whole-mount in situ hybridization
- WT:
-
Wild-type
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Funding
The research conducted in this manuscript was funded by the National Natural Science Foundation of China (82000458) to CC and start-up funding for Doctoral Research of Nantong Science and Technology College (NTKY-Dr2017001) to FL. It is also funded by Nantong Science and Technology Program (JC2021088), Open Program of Key Laboratory of Cultivation and High-value Utilization of Marine Organisms in Fujian Province (2019fjsccq08), Natural Science Foundation of the Jiangsu Higher Education Institutions of China (20KJB180008), and the Science Foundation of Nantong City (JC2020023).
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CC and DL designed and conceptualized the project. FL, XG, PQ, and XL performed the experiment. FL and CC analyzed the data. CC wrote the manuscript. All authors read and approved the final manuscript.
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All animal-related experiments were carried out following the NIH Guidelines for the care and use of laboratory animals (http://oacu.od.nih.gov/regs/index.htm), and animal protocols were ethically approved by the Administration Committee of Experimental Animals of Nantong University, Jiangsu Province, China (Approval ID: 20180608-Z001). Transgenic zebrafish lines provided by others were approved by the owners with written informed consent.
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Feng Lv, Xiaojuan Ge and Peipei Qian are contributed equally to this work.
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10695_2022_1049_MOESM1_ESM.png
Supplementary Figure S1. Expressions of cardiac-related genes in WT and nav3-/- mutants at 24 hpf. The expression of the corresponding gene is outlined with a white dashed line. The digits at the lower left position of each panel indicate the number of fishes with typical phenotype in total observed ones. (PNG 3028 KB)
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Lv, F., Ge, X., Qian, P. et al. Neuron navigator 3 (NAV3) is required for heart development in zebrafish. Fish Physiol Biochem 48, 173–183 (2022). https://doi.org/10.1007/s10695-022-01049-5
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DOI: https://doi.org/10.1007/s10695-022-01049-5